| Objective:Adopting the method of MCAO(middle cerebral artery occlusion) to make model,es-tablishing the rats infarction model,to observe the effect of Pien Tze Huang on infla-mmatory injury related factors of IL-1β,IL-6,TNF-α,blood brain barrier factors of Z O-1,MMP-9and neurotrophic factor of BDNF,from the protein level and gene level to discuss Pien Tze Huang in the treatment of cerebral infarction efficacy and mechanis-ms,elucidating Pien Tze Huang having neuroprotective effect.Methods:Adult health male SD rats,weight260-280g.Rats were randomly divided into operati-on and non-operation group(normal group).The operation group applied improved Lon-ga method to make the focal and permarent model of MCAO,and implemented EZ-L-onga evaluation after the rats woke up for2hours.According to the evaluation the ra-ts included in the standards were randomly divided into model group,Pien Tze Huang high-dose group,Pien Tze Huang middle-dose group,Pien Tze Huang small-dose group, nimodiping group,naodesheng group and normal saline group.The above.groups were ad ministered for3days before making model,on the fourth day making the MCAO,on t he seventh day were observed.after anaesthesia in rats,through the abdominal aortabloo-d.serum for ELISA detection IL-1β,IL-6,TNF-α expression changes;and then took the brain tissue depending on the cross2mm before and after to dehydrate,make paraffin section.Quickly brung out infarction brain tissue about180-200mg in liquid nitrogen to preserve,finally the expression changes of ZO-1,MMP-9would be observed by mothed-s of western blot and RT-PCR.Results:1.MCAO model preparation and evaluation.In this study,postoperative neurologic scores in MCAO were evaluated.No neurologi-cal deficit symptoms of the normal group,score O.model group scores were significant-1y higher than the normal group,severe symptoms of neurological deficit,suggesting thatt he model was successful.2.Using immunohistochemistry to detect middle cerebral artery occlusion induced focal cerebral ischemia rats neurotrophic factor BDNF,and inflammatory injury factor IL-6expression.In this study,using immunohistochemistry method to observe different regions(Infarcti on surrounding area, in the central area of infarction area,infarction of the contralateral normal area)of BDNF variation,the result shows:infarction of the surrounding area:them odel group compared with the sham group was significantly increased(P<0.05),treatmen t group expression of BDNF were higher than the model group,which Pien Tze Huan g high dose group,Pien Tze Huang middle dose group and nimodipine group than mo del group differences were significant(P<0.01);Pien Tze Huang high dose group and P-ien Tze Huang small dose group have a significant difference(P<0.05).Infarction of the central area:The BDNF expression of model group was significantly lower than the n-ormal group(P<0.01),the treatment group were higher than the model group,naodesheng group,nimodipine group and Pien Tze Huang high dose group have a significant diffe-rence than model group differences(P<0.01or P<0.05).Normal zone:model group,the e-xpression of BDNF increased than the normal group,the treated group were higher th-an the model group,no ignificant differences among hese groups(P>0.05).In this study,using immunohistochemistry method to observe different regions(lnfarcti on surrounding area, in the central area of infarction area,infarction of the contral ater al normal area) of IL-6variation,the result showsrinfarction of the surrounding area:the model group compared with the sham group was significantly increased(P<0.05),treat ment group expression of IL-6were lower than the model group,which Pien Tze Hua ng high dose group,naodesheng group and nimodipine group compared with model gro up differences were significant(P<0.01);Pien Tze Huang high dose group and Pien Tze Huang small dose group have a significant difference(P<0.05).Infarction of thecentral area:The IL-6expression of model group was significantly lower than the normal grou p(P<0.01),the treatment group were higher than the model group,naode-sheng group,ni modipine group and Pien Tze Huang high dose group have a significa-nt difference th an model group differences(P<0.01or P<0.05).Normal zone:model group,the expression of IL-6increased than the normal group,the treated group werelower than the model group,no significant differences among these groups (P>0.05).3.Using ELISA methord to detect middle cerebral artery occlusion induced focal cerebral ischemia rats inflammatory injury related factors IL-1β,IL-6and TNF-α.Detected by ELISA methord in MCAO rats,the expression of IL-1β, the results show: the model group was significantly increased(P<0.01)compared with the sham group,Pie n Tze Huang high dose group,Pien Tze Huang middle dose group,Pien Tze Huang sm all dose group,naodesheng group and nomodiping group of IL-1β expressionwere signif icantly reduced compared with the model group(P<0.01or P<0.05).Detected by ELISA methord in MCAO rats,the expression of IL-6,the results show:t he model group was significantly increased compared with the sham group(P<0.01),Pie n Tze Huang high dose group,naodesheng group and nomodiping group of IL-6expres sion were significantly reduced compared with the model group(P<0.01or P<0.05).Detected by ELISA methord in MCAO rats,the expression of TNF-a,the results sho w:the model group was significantly increased(P<0.01) compared with the sham group, Pien Tze Huang high dose group,Pien Tze Huang middle dose group,Pien Tze Huang small-dose group,naodesheng group and nomodiping group of TNF-a expression were s ignificantly reduced compared with the model group(P<0.01or P<0.05).4.Using RQ-PCR methord to detect middle cerebral artery occlusion induced focal c-erebral ischemia rats blood-brain barrier related factors ZO-1,MMP-9mRNA.The results show that ZO-l:the model group compared with the sham group was si-gnificantly lower(P<0.05).treated groups of ZO-1expression were higher than the mode1group,which Pien Tze Huang high dose group,Pien Tze Huang middle dose group an d nimodipine group were significant(P<0.01);Pien Tze Huang high dose group compare d with Pien Tze Huang low-dose group had significant difference(P<0.05).The results show that MMP-9:the model group compared with the sham group was significantly higher(P<0.05).treated groups of MMP-9expression were lower than the model group,which Pien Tze Huang high dose group,naodesheng group and nimodipine group were significant(P<0.01);Pien Tze Huang high-dose group compared with Pien Tze Huang low-dose group had significant difference(P<0.05).5.Using Western blot method detect middle cerebral artery occlusion induced focal c erebral ischemia rats blood-brain barrier related factors ZO-1,MMP-9protein.The results show that ZO-l:the model group compared with the sham group was si-gnificantly lower(P<0.05).treated groups of ZO-1expression were higher than the mod el group,which Pien Tze Huang high dose group,Pien Tze Huang middle dose group a nd nimodipine group were significant(P<0.01);Pien Tze Huang high-dose group compar ed with Pien Tze Huang low-dose group had significant difference(P<0.05).The results show that MMP-9:the model group compared with the sham group was significantly higher(P<0.05).treated groups of MMP-9expression were lower than the model group,which Pien Tze Huang high dose group,naodesheng group and nimodi-pin e group were significant(P<0.01);Pien Tze Huang high-dose group compared with Pien Tze Huang low-dose group had significant difference(P<0.05).Conclusion:1Pien Tze Huang can promote BDNF,reduce IL-6protein level after cerebral ischemia;2Pien Tze Huang can reduce cerebral infarction of serum IL-1β,IL-6and TNF-α content;3Pien Tze Huang can promote cerebral infarction of ZO-1,reduce the expression levels of MMP-9mRNA;4Pien Tze Huang can promote cerebral infarction of ZO-1,reduce the level of MMP-9protein expression;5Pien Tze Huang has the function of cerebral protection,inhibiting the inflammatory injury,improving the blood-brain barrier mechanism after cerebral ischemia. |
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